Inhalation of high concentrations of hydrogen ameliorates liver ischemia/reperfusion injury through A2A receptor mediated PI3K-Akt pathway
Graphical abstract
Introduction
Hepatic ischemia and reperfusion (I/R) injury is a phenomenon in which cellular damage is induced by hypoxia following the restoration of blood flow and oxygen delivery after transplantation surgery, tissue resections, and hemorrhagic shock [1]. Pathologically, liver I/R injury may cause hepatocyte swelling, hepatocyte vacuolization, endothelial cell disruption, neutrophil infiltration, and hepatocyte necrosis and apoptosis [2]. Hepatic I/R injury may significantly compromise graft survival and postoperative liver function, resulting in a high mortality. It affects liver function and significantly increases the risk to the circulatory system and respiratory system [3]. To date, numerous studies have explored the treatment and prevention of liver I/R injury [4], [5], [6], but no effective strategies have been developed. Therefore, liver I/R injury is an important clinical problem that requires further study.
In the pathogenesis of hepatic I/R injury, oxidative stress and inflammation are the two major mechanisms, and some strategies targeting reactive oxygen species (ROS) and inflammation are used for the treatment of hepatic I/R injury [7], [8].
Hydrogen is the simplest molecule in nature. It not only exists in nature but also can be generated in the human intestine. Traditionally, it is believed to function as an inert gas at body temperature in mammalian cells because it cannot react with biological compounds, including oxygen (O2) gas, in the absence of catalysts at body temperature. Thus, hydrogen has been used during deep diving for the prevention of nitrogen narcosis [9]. In recent years, hydrogen has been found to protect against I/R injury to the brain [10], heart [11], kidney [12], liver [13], [14], and retina [15], mainly by scavenging hydroxyl radicals, inhibiting inflammation, and suppressing cell apoptosis. However, in most studies, 2% or 4% hydrogen gas was used [10]. Recently, our group treated diseases with high concentrations of hydrogen (HCH) gas (67% H2, 33% O2) in animal models [15], [16]. The mixed gas is produced using an AMS-H-01 hydrogen oxygen nebulizer (Asclepius, Shanghai, China), which can produce H2 and O2 by electrolyzing water [15], [16]. Whether HCH is also protective towards hepatic I/R injury and, if so, the mechanism underlying the hepatoprotection of HCH remains unclear.
The phosphatidylinositol-3-kinase (PI3K)/Akt pathway is important for cell survival, and activation of the PI3K/Akt pathway has been found to protect cells against injury. Previous studies have shown that PI3K/Akt pathway activation is important for protection against I/R injury [17], [18]. In addition, studies have shown that A2A receptor is involved in hepatic protective effect [19], and the PI3K/Akt pathway can be regulated by A2A receptor [20].
This study was performed to explore the protective effects of HCH on hepatic I/R injury and to examine the role of the A2A receptor and PI3K/Akt pathway in the protective effects of HCH.
Section snippets
Animals
A total of 96 male C57BL/6 wild-type (WT) mice aged 8–10 weeks and weighing 20–25 g were purchased from the Experimental Animal Center of the Second Military Medical University, Shanghai, China. Mice were housed in a specific pathogen free environment with a 12-h/12-h light/dark cycle and given ad libitum access to food and water. All procedures were performed according to the recommendations of the Committee of the Care and Use of Laboratory Animals at the Second Military Medical University.
Inhibition of the PI3K/Akt pathway activation attenuates the hepatoprotective effects of HCH on hepatic I/R injury
Serum ALT in the I/R group (8351.583 ± 1310.280 IU/L) increased significantly compared with the Sham group (25.833 ± 8.747 IU/L) as shown in Fig. 2A. HCH significantly decreased I/R induced increases in serum ALT (1739.583 ± 419.826 IU/L). LY294002, an inhibitor of PI3K, significantly increased serum ALT after HCH treatment (7099.417 ± 973.170 IU/L). Moreover, there was a significant difference in the serum ALT between I/R group and LY + I/R + HCH group. However, there was no significant difference in serum
Discussion
In this study, we investigated the hepatoprotective effects of HCH in a mouse model. Our results showed that, after liver I/R injury, inhalation of hydrogen at a high concentration (66.7% H2) improved the liver pathology and liver function, reduced the oxidative stress and inflammation in the liver, and inhibited the apoptosis of hepatocytes after I/R injury, which were, at least partially, related to the activation of A2A receptor mediated PI3K/Akt pathway because inhibition of this pathway
Conflict of interest
There is no conflict of interest in this paper.
Author contribution statement
Li H., Che O. and Liu W. wrote the main manuscript. Ye Z. prepared Fig. 1, Fig. 2. Li H. finished Fig. 5, Fig. 6. Sun X. designed all figures. Zhang R. revised the whole manuscript. Zhang N. finished Fig. 3, Fig. 4. Huang J. designed the supplements 1.
Acknowledgements
This study was supported by the National Natural Science Foundation of China – China (No. 81371316). The English in this document has been checked by at least two professional editors, both native speakers of English. For a certificate, please see: http://www.textcheck.com/certificate/4TMRNL
References (39)
- et al.
Hepatic warm ischemia-reperfusion-induced increase in pulmonary capillary filtration is ameliorated by administration of a multidrug resistance-associated protein 1 inhibitor and leukotriene D4 antagonist (MK-571) through reducing neutrophil infiltration and pulmonary inflammation and oxidative stress in rats
Transplant. Proc.
(2015) - et al.
Current strategies to minimize hepatic ischemia-reperfusion injury by targeting reactive oxygen species
Transplant. Rev. (Orlando)
(2012) - et al.
Inhalation of hydrogen gas suppresses hepatic injury caused by ischemia/reperfusion through reducing oxidative stress
Biochem. Biophys. Res. Commun.
(2007) - et al.
Postconditioning with inhaled hydrogen promotes survival of retinal ganglion cells in a rat model of retinal ischemia/reperfusion injury
Brain Res.
(2016) - et al.
Role of phosphatidylinositol 3-kinase in the development of hepatocyte preconditioning
Gastroenterology
(2004) - et al.
Helium preconditioning protects mouse liver against ischemia and reperfusion injury through the PI3K/Akt pathway
J. Hepatol.
(2014) - et al.
FGL2/fibroleukin mediates hepatic reperfusion injury by induction of sinusoidal endothelial cell and hepatocyte apoptosis in mice
J. Hepatol.
(2012) - et al.
Methane attenuates myocardial ischemia injury in rats through anti-oxidative, anti-apoptotic and anti-inflammatory actions
Free Radic. Biol. Med.
(2016) - et al.
JNK mediates hepatic ischemia reperfusion injury
J. Hepatol.
(2005) - et al.
Inhibition of Hedgehog signaling for the treatment of lymphoma and CLL: a phase II study from the LYSA
Ann. Oncol.
(2016)
Stat4 and Stat6 signaling in hepatic ischemia/reperfusion injury in mice: HO-1 dependence of Stat4 disruption-mediated cytoprotection
Hepatology
The protective of hydrogen on stress-induced gastric ulceration
Int. Immunopharmacol.
Pharmacological inhibitors of glycogen synthase kinase 3
Trends Pharmacol. Sci.
Regulation of mTORC1 by PI3K signaling
Trends Cell Biol.
Molecular mechanisms of hepatic ischemia-reperfusion injury and preconditioning
Am. J. Physiol. Gastrointest. Liver Physiol.
Ischemia/reperfusion injury in liver surgery and transplantation: pathophysiology
HPB Surg.
Preconditioning methods in the management of hepatic ischemia reperfusion- induced injury: update on molecular and future perspectives
Hepatol. Res.
A systematic review of pharmacological treatment options used to reduce ischemia reperfusion injury in rat liver transplantation
PLoS One
The mechanisms and strategies to protect from hepatic ischemia-reperfusion injury
Eur. Rev. Med. Pharmacol. Sci.
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Both contributed to this paper equally.